Rat Serum Albumin Research Articles

Quantitative cross-species comparison of serum albumin binding of per- and polyfluoroalkyl substances from five structural classes.

Per- and polyfluoroalkyl substances (PFAS) are a class of over 8000 chemicals, many of which are persistent, bioaccumulative, and toxic to humans, livestock, and wildlife. Serum protein binding affinity is instrumental in understanding PFAS toxicity, yet experimental binding data is limited to only a few PFAS congeners. Previously, we demonstrated the usefulness of a high-throughput, in vitro differential scanning fluorimetry assay for determination of relative binding affinities of human serum albumin for 24 PFAS congeners from 6 chemical classes. In the current study, we used this assay to comparatively examine differences in human, bovine, porcine, and rat serum albumin binding of 8 structurally informative PFAS congeners from 5 chemical classes. With the exception of the fluorotelomer alcohol 1H, 1H, 2H, 2H-perfluorooctanol (6:2 FTOH), each PFAS congener bound by human serum albumin was also bound by bovine, porcine, and rat serum albumin. The critical role of the charged functional headgroup in albumin binding was supported by the inability of albumin of each species tested to bind 6:2 FTOH. Significant interspecies differences in serum albumin binding affinities were identified for each of the bound PFAS congeners. Relative to human albumin, perfluoroalkyl carboxylic and sulfonic acids were bound with greater affinity by porcine and rat serum albumin, and the perfluoroalkyl ether acid congener bound with lower affinity to porcine and bovine serum albumin. These comparative affinity data for PFAS binding by serum albumin from human, experimental model, and livestock species reduce critical interspecies uncertainty and improve accuracy of predictive bioaccumulation and toxicity assessments for PFAS.

Elucidating the mechanism of the Tibetan medicine sanguotang in treating gouty arthritis through network pharmacology and in vivo experiments

BackgroundWe explored the mechanisms of Sanguotang (SGT), a Tibetan medicine, in treating gout arthritis (GA). MethodsThe main active components, action targets, and disease targets of SGT were identified through TCMSP databases. The gene functions were analyzed using protein interaction (PPI) networks, Gene Ontology (GO) enrichment analysis, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis, and molecular docking. A GA model induced by monosodium urate was established in rats. The ankle joint swelling was observed. The levels of uric acid (UA) and albumin (ALB) in rat serum were measured. Hematoxylin and eosin (HE) staining was conducted to examine the pathological changes in rat ankle joints. ResultsTwenty-nine active components of SGT with proven efficacy and 66 intersection targets were identified, primarily involved in inflammation and immune regulation pathways. The PPI results revealed that the key targets of SGT against GA included ALB, IL6, TNF, TP53, and PTGS. Molecular docking showed favorable binding energy between the ALB protein and the active components. The results from animal experiments demonstrated that SGT effectively alleviated the inflammatory reaction in ankle joints, and decreased UA and ALB levels. Furthermore, SGT effectively inhibited the proliferation of synovial cells in the ankle joint cavity, prevented infiltration of inflammatory cells, and protected synovial tissue, thereby improving GA. ConclusionsSGT comprehensively contributes to the treatment of GA by regulating UA metabolism, reducing the release of inflammatory factors, and modulating immune and inflammatory pathways.

Functionalisation of brush polyethylene glycol polymers with specific lipids extends their elimination half-life through association with natural lipid trafficking pathways

Polymeric prodrugs have been applied to control the delivery of various types of therapeutics. Similarly, conjugation of peptide therapeutics to lipids has been used to prolong systemic exposure. Here, we extend on these two approaches by conjugating brush polyethylene glycol (PEG) polymers with different lipid components including short-chain (1C2) or medium-chain (1C12) monoalkyl hydrocarbon tails, cholesterol (Cho), and diacylglycerols composed of two medium-chain (2C12) or long-chain (2C18) fatty acids. We uniquely evaluate the integration of these lipid-polymers into endogenous lipid trafficking pathways (albumin and lipoproteins) and the impact of lipid conjugation on plasma pharmacokinetics after intravenous (IV) and subcutaneous (SC) dosing to cannulated rats. The IV and SC elimination half-lives of Cho-PEG (13 and 22 h, respectively), 2C12-PEG (11 and 17 h, respectively) and 2C18-PEG (12 h for both) were prolonged compared to 1C2-PEG (3 h for both) and 1C12-PEG (4 h for both). Interestingly, 1C2-PEG and 1C12-PEG had higher SC bioavailability (40 % and 52 %, respectively) compared to Cho-PEG, 2C12-PEG and 2C18-PEG (25 %, 24 % and 23 %, respectively). These differences in pharmacokinetics may be explained by the different association patterns of the polymers with rat serum albumin (RSA), bovine serum albumin (BSA) and lipoproteins. For example, in pooled plasma (from IV pharmacokinetic studies), 2C18-PEG had the highest recovery in the high-density lipoprotein (HDL) fraction. In conclusion, the pharmacokinetics of brush PEG polymers can be tuned via conjugation with different lipids, which can be utilised to tune the elimination half-life, biodistribution and effect of therapeutics for a range of medical applications. Statement of significanceLipidation of therapeutics such as peptides has been employed to extend their plasma half-life by promoting binding to serum albumin, providing protection against rapid clearance. Here we design and evaluate innovative biomaterials consisting of brush polyethylene glycol polymers conjugated with different lipids. Importantly, we show for the first time that lipidated polymeric materials associate with endogenous lipoprotein trafficking pathways and this, in addition to albumin binding, controls their plasma pharmacokinetics. We find that conjugation to dialkyl lipids and cholesterol leads to higher association with lipid trafficking pathways, and more sustained plasma exposure, compared to conjugation to short and monoalkyl lipids. Our lipidated polymers can thus be utilised as delivery platforms to tune the plasma half-life of various pharmaceuticals.

(Pro)Renin Receptor Decoy Peptide PRO20 Protects against Oxidative Renal Damage Induced by Advanced Oxidation Protein Products

Chronic kidney disease (CKD) is associated with advanced oxidation protein products (AOPPs). A recent study has shown that AOPP-induced renal tubular injury is mediated by the (pro)renin receptor (PRR). However, it is unclear whether the PRR decoy inhibitor PRO20 can protect against renal damage related to AOPPs in vivo. In this study, we examined the role of the PRR in rats with AOPP-induced renal oxidative damage. Male SD rats were subjected to unilateral nephrectomy, and after a four-day recuperation period, they were randomly divided into four groups (n = 6/group) for four weeks: control (CTR), unmodified rat serum albumin (RSA, 50 mg/kg/day via tail-vein injection), AOPPs-RSA (50 mg/kg/day via tail-vein injection), and AOPPs-RSA + PRO20 (50 mg/kg/day via tail-vein injection + 500 μg/kg/day via subcutaneous injection) groups. PRO20 was administered 3 days before AOPPs-RSA injection. Renal histopathology evaluation was performed by periodic acid-Schiff (PAS) staining, and biochemical parameters related to renal injury and oxidative stress biomarkers were evaluated. The expression of related indicators was quantified by RT-qPCR and immunoblotting analysis. In the results, rats in the AOPPs-RSA group exhibited higher levels of albuminuria, inflammatory cell infiltration, and tubular dilation, along with upregulation of oxidative stress, profibrotic and proinflammatory factors, and elevation of AOPP levels. Meanwhile, in the PRO20 group, these were significantly reduced. Moreover, the levels of almost all components of the renin-angiotensin system (RAS) and Nox4-dependent H2O2 production in urine and the kidneys were elevated by AOPPs-RSA, while they were suppressed by PRO20. Furthermore, AOPPs-RSA rats showed elevated kidney expression of the PRR and soluble PRR (sPRR) and increased renal excretion of sPRR. In summary, these findings suggest that PRR inhibition may serve as a protective mechanism against AOPP-induced nephropathy by inhibiting the intrarenal RAS and Nox4-derived H2O2 mechanisms.

Evaluation of Acute Effects of Cadmium Toxicity on Serum Total Protein, Albumin and Globulin in Male Albino Wistar Rats Treated with Ethanol leaf Extracts of Tapinanthus bangwensis and Mangifera indica

Acute impacts of heavy metal exposure on serum total protein (TP), albumin (AL) and globulin (GL) in male albino Wistar rats treated with ethanol leaf extracts of Tapinanthus bangwensis and Mangifera indica were evaluated. Twenty-five (25) animals (125-250 g) were randomly assigned five groups of five rats each. Groups 1-4 were orally administered with cadmium chloride (30 %) below its LD50, (267 mg/kg) each for three times at 2 days interval before treatment with the plant extracts. Groups 1 and 2 were later treated with 300 mg/kg of T. bangwensis and M. indica respectively. Group 3 was treated with 300 mg/kg combined extracts of the two plants at 50:50 dosage ratio while group 4 and 5 were not treated with the plant extracts and served as positive and normal control respectively. Animals in each group were allowed free access to commercial rat mash and water throughput two weeks of treatment which was done daily via oral route. After the fourteen (14) days experimentation, the results revealed a significant increase (P<0.05) in serum total protein in group 2 and 3 when compared to group 1. No significant difference (P>0.05) was observed in group 1 compared to the positive and normal controls. Group 2 showed significant increase (P<0.05) when compared to groups 3 and 5. Group 3 showed significant increase (P<0.05) when compared to groups 4 and 5, but with no significant difference (P>0.05) in group 4 compared to group 5. Serum albumin level showed significant increase (P<0.05) in group 1 when compared to group 2 and 3, but with no significant difference (P>0.05) compared to groups 4 and 5. Significant decrease in group 2 (P<0.05) when compared to group 4. Group 4 compared to group 5. Significant increase in serum albumin was revealed in group 4 when compared to group 5. Serum globulin recorded significant decrease (P<0.05) in group 1 compared to group 2 and 3, but with no significant difference (P>0.05) compared to group 4 and 5. Group 2 showed significant increase (P<0.05) when compared to groups 4 and 5. Group 3 recorded significant increase (P<0.05) in serum globulin when compared to Group 4, but with no significant difference (P>0.05) compared to group 5. The implications of these results are that human should not be exposed to heavy metals to avid toxicity, also if exposed the extracts used could be administered to ameliorate.

Effects of amino acids and albumin administration on albumin metabolism in surgically stressed rats: A basic nutritional study.

Nutrition therapy and administration of albumin preparations are common in postsurgical patients. However, the effects of these interventions on albumin metabolism are unclear. We elucidated the effect of postoperative albumin and/or parenteral nutrition administration on it. Sprague-Dawley rats underwent surgery involving intestinal rubbing followed by intestinal exposure. Subsequently, they were administered experimental solutions for 48 h, their blood samples were collected at 24 and 48 h, and livers were excised at 48 h. Based on experimental solutions, rats were divided into five groups: non-surgical (Non-surg); glucose and electrolyte solution (GE); amino acid, glucose, and electrolyte solution (AGE); GE + rat serum albumin (Alb) (GE + Alb); and AGE + Alb. Their plasma albumin concentrations; albumin fractional synthesis rate (ALB FSR); mercaptoalbumin/total albumin ratio (MA ratio); and messenger RNA (mRNA) expressions of albumin and hepatocyte nuclear factor-1 (HNF-1) in the liver were measured. The GE and AGE groups showed significant decline in albumin concentrations. ALB FSR was significantly enhanced in the AGE group compared with the GE group. The mRNA expression of albumin was similar to ALB FSR in all groups and that of HNF-1 was significantly decreased in the GE + Alb and AGE + Alb groups compared with the Non-surg group. The MA ratio in the AGE group was similar to the Non-surg group. The administration of amino acids comprising parenteral nutrition after surgery augmented ALB FSR and maintained the MA ratio only without simultaneous albumin administration.

Effect of Saccharum barberi on the total serum protein and serum albumin in Albino wistar rats and Guinea pigs

Effect of Saccharum barberi on the total serum protein and serum albumin in Albino wistar rats and Guinea pigs

333.9: The Effect of Short-term Inhalation of Isoflurane on the Outcomes of Intraportal Hepatocyte Transplantation

Introduction: Liver transplantation is currently established as a treatment for endo-stage liver disease. However, this treatment is considered too invasive for those patients. Hepatocyte transplantation (HTx), which is less invasive in comparison to liver transplantation, has been expected to serve as an alternative therapy to liver transplantation. However, the outcomes of HTx are still far from satisfactory, and it is well known that there is a large discrepancy between the experimental results of animal models and clinical outcomes of HTx. Of particular note, clinical HTx is only performed without general anesthesia, while inhalation anesthetics are usually used in animal experiments. Isoflurane, which is one of the most widely used inhalation anesthetics in animal experiments, is known to have a strong vasodilatory effect, portal pressure inhibitory effect, and cytoprotective effect against cytokine-induced injury. Therefore, we hypothesized that isoflurane may be a possible reason for the discrepancy between the results of animal experiments and the clinical outcomes of HTx. Methods: F344/NSLc rat hepatocytes were isolated. Isolated hepatocytes (1.0×107 cells) were transplanted to analbuminemic rats with (ISO group) and without (AW group) isoflurane. These analbuminemic rats had a syngeneic background to the donor rats. The serum albumin, AST, ALT, LDH levels and several inflammatory mediators in the recipient rats were analyzed. Immunohistochemical staining and ex vivo imaging of transplanted hepatocytes were also performed. Results: The serum albumin levels of the ISO group were significantly higher in comparison to the AW group (p<0.05) (Figure 1). The serum AST, ALT, LDH levels of the ISO group were significantly suppressed in comparison to the AW group (p<0.0001, respectively). The serum levels of IL-1β (p<0.01), IL-10 (p<0.01), IL-18 (p<0.01), MCP-1 (p<0.05), RANTES (p<0.01), Fractalkine (p<0.01), and LIX (p<0.05) in the ISO group were significantly downregulated in comparison to the AW group. The ischemic regions of the recipient livers, which were easily detected in the livers of both groups, tended to be more evident in the AW group (3.69±1.05%) in comparison to the ISO group (2.42±1.76%) (Figure 2). Conclusions: This study showed that the anti-inflammatory effects of isoflurane could efficiently contribute to hepatocyte engraftment. According to the detailed analyses in this study, the anti-inflammatory effects, rather than vasodilatory effects, of isoflurane appeared to be the main mechanism of the abovementioned benefits. Isoflurane may at least in part be a reason for the discrepancy between the results of animal experiments and the clinical outcomes of HTx.

Naringenin antihypertensive power is mediated via abrogation of mineralocorticoid receptor (MCR)/ angiotensin converting enzyme (ACE)/ kidney injury molecule (Kim‐1) signaling pathway

Hypertension is a condition with chronic elevation of blood pressure and a common preventable risk factor for cardiovascular disease with attendant global morbidity and mortality. The present study investigated the novel antihypertensive action of Naringenin on NƱ‐Nitro‐L‐arginine Methyl Ester (L‐NAME)‐induced hypertension together with possible molecular mechanism of action. Rats were divided into four groups. Rats in Group A were normotensive. The hypertensive group (Group B) received 40 mg/kg) of L‐NAME alone while Groups C and D were concurrently administered Naringenin (50 mg/kg) or Lisinopril (10 mg/Kg) together with L‐NAME orally for 3 weeks. Blood pressure parameters, markers of oxidative stress and renal damage were measured. The immunohistochemistry of kidney injury molecule 1, mineralocorticoid receptor and angiotensin converting enzyme were also determined. Results indicated significant increase in oxidative stress markers and decrease in serum nitric oxide bioavailability in hypertensive rats. Furthermore, there were significant increases in serum myeloperoxidase activity, urinary creatinine, albumin, and blood urea nitrogen levels in hypertensive rats in comparison to hypertensive rats treated with Naringenin. Immunohistochemistry revealed higher expression of kidney injury molecule 1, mineralocorticoid receptor and angiotensin converting enzyme in hypertensive rats. However, co‐treatment with Naringenin mitigated renal oxidative stress, normalized blood pressure and lowered the expression of kidney injury molecule 1, mineralocorticoid receptor, and angiotensin converting enzyme. Combining all, Naringenin offered a novel antihypertensive action through the down regulation of kidney injury molecule 1, deactivation of mineralocorticoid receptor and inhibition of angiotensin converting enzyme.

Absorption, Distribution, Metabolism, and Excretion of [14C]-Sotorasib in Rats and Dogs: Interspecies Differences in Absorption, Protein Conjugation and Metabolism.

Sotorasib is a first-in-class, targeted covalent inhibitor of Kirsten rat sarcoma viral oncogene homolog (KRAS)G12C approved by the FDA to treat patients with locally advanced or metastatic non-small cell lung cancer with the KRASG12C mutation. The mass balance, excretion, and metabolism of [14C]-sotorasib was characterized in rats and dogs after a single dose of 60 or 500 mg/kg, respectively. Mean recovery was >90% for both species. Excretion of unchanged sotorasib was a minor pathway in rats, accounting for <4% of administered dose in urine and <7% of administered dose in feces. Approximately 66% of administered dose was recovered in the bile from bile duct cannulated rats as metabolites. Excretion of unchanged sotorasib was the major excretion pathway in dogs, likely caused by solubility-limited absorption. Major pathways of sotorasib biotransformation included glutathione conjugation and oxidative metabolism. In vitro experiments demonstrated that nonenzymatic conjugation (Michael addition) was the primary mechanism of the reaction with glutathione. Extended radioactivity profiles in blood and plasma were observed in rats, but not dogs, after dosing with [14C]-sotorasib. In vitro experiments demonstrated that sotorasib-protein adducts were observed with both rat hemoglobin and serum albumin, explaining the extended radioactivity profile. SIGNIFICANCE STATEMENT: This study characterized the mass balance, excretion, and metabolism of [14C]-sotorasib, a covalent Kirsten rat sarcoma viral oncogene homolog G12C inhibitor, in rats and dogs. Rapid absorption and extensive metabolism of sotorasib was observed in rats, while sotorasib was primarily excreted unchanged in dog feces, likely due to solubility-limited absorption. Protein adducts with rat hemoglobin and serum albumin were characterized, explaining observed extended blood and plasma radioactivity profiles. The primary biotransformation pathway, glutathione conjugation, was mediated through nonenzymatic conjugation.

Yi Shen An, a Chinese traditional prescription, ameliorates membranous glomerulonephritis induced by cationic bovine serum albumin in rats

Context Yi Shen An (YSA) is an investigational composite of traditional Chinese medicine (Reference: 2010L000974) for the treatment of renal disease. Objective To investigate the protective effects of YSA against membranous glomerulonephritis (MGN). Materials and methods Male Sprague–Dawley rats were injected with cationic bovine serum albumin (C-BSA) to create a model of MGN. Then, rats were orally treated with YSA at doses of 0.25, 0.5, 1 and 2 g/kg for 35 successive days; prednisone (5 mg/kg) was used as a positive control. At the end of the experimental period, we performed a series of tests, including 24 h urinary protein, and biochemical, immunological, antioxidative, coagulation indices, and histopathological examination. Results YSA-1 g/kg significantly lowered urinary protein from 68.37 to 30.74 mg (p < 0.01). Meantime, total protein (TP) and albumin (ALB) recovered from 66.26 and 20.51 g/L to 76.08 and 35.64 g/L (p < 0.01), respectively. YSA removed the deposition of immunoglobulin G (IgG) and complement 3c (C3c), prevented inter-capillary cell hyperplasia on the glomerular basement membrane (GBM), and reduced electron-dense deposits and fusion of podocytes. In addition, serum IgG and superoxide dismutase were significantly elevated. In contrast, malondialdehyde, total cholesterol, triglyceride, circulating immune complex (CIC), and immunoglobulin M decreased in the YSA-treated group. Moreover, the blood coagulation dysfunction was adjusted. Discussion and conclusions These findings indicate YSA may exert a therapeutic effect against MGN through the inhibition of CIC formation, and the removal of IgG and C3c deposition from the GBM, thus supporting the development of further clinical trials.

Formulation and evaluation of the vascular endothelial growth factor loaded polycaprolactone nanoparticles

In an attempt to increase molecular stability and provide controlled release, vascular endothelial growth factor (VEGF) was encapsulated into polycaprolactone (PCL) nanoparticles. Both VEGF-free and VEGF-loaded PCL nanoparticles were formulated by w/o/w double emulsion of the dichloromethane-water system in the presence of polyvinyl alcohol (PVA) and rat serum albumin. To achieve the optimal formulation concerning particle size and monodispersity, studies were carried out with different formulation parameters, including PVA concentration, homogenization time and rate. Scanning electron microscopy and dynamic light scattering analysis showed respectively that particles had a spherical shape with a smooth surface and particle size varying between 58.68-751.9 nm. All of the formulations were negatively charged according to zeta potential analysis. In vitro release study was performed in pH 7.4 phosphate-buffered saline at 37°C and released VEGF amount was measured by enzyme-linked immunosorbent assay (ELISA) method. At the end of the 35th day, 10% of total encapsulated VEGF was released with a sustained-release profile, which fitted the Korsmeyer-Peppas kinetic model. The bioactivation of the nanoparticles was evaluated using XTT and ELISA methods. As a result, the released VEGF was biologically active and also VEGF loaded PCL nanoparticles enhanced proliferation of the human umbilical vein endothelial cells in cell culture.

Doxorubicin alters the disposition of phenytoin by reducing its metabolic elimination and binding affinity to serum albumin in rats.

We investigated the pharmacokinetic interaction of doxorubicin (DOX) with phenytoin (PHT) and the underlying mechanism in rats to clarify why the serum PHT concentration decreases despite the impaired PHT metabolic capacity in patients receiving DOX. Rats were administered 15 mg/kg of DOX or saline alone. The pharmacokinetic disposition of intravenously administered PHT was examined 4 days after DOX exposure. Enzyme kinetics of CYP2C-dependent PHT p-hydroxylation were analysed using hepatic microsomes. The unbound PHT concentration in serum was measured by the ultrafiltration method, and the relationship between the unbound fraction (fu) and serum albumin level was assessed. The total clearance (CLtot) of PHT was significantly increased by DOX, but the activity of PHT p-hydroxylation conversely decreased. The unbound serum PHT concentration and its fu were significantly higher in the DOX group than in the control group, and the CLtot/fu, a measure of intrinsic clearance, significantly decreased. An increase in the fu was observed even when using a serum sample with an albumin concentration equal to that in the control group. DOX treatment increases the unbound serum PHT concentration by depressing the metabolic capacity and alters the total PHT by reducing serum albumin and its affinity to PHT.

Nephroprotective activity of Vetiveria zizanioides (L.) Nash supplement in doxorubicin-induced nephrotoxicity model of Wistar rats.

The nephroprotective effect of standardized aqueous root extract of Vetiveria zizanioides (L.) Nash (Family: Poaceae) was investigated in doxorubicin-induced (20mg/kg, ip) experimental nephrotoxicity model of Wistar rats. The freeze-dried aqueous refluxed (4hr) root extract of V. zizanioides (25, 50; equivalent human therapeutic dose and 100mg/kg) was administered separately to nephrotoxic Wistar rats (n=6/group). Supplement of V. zizanioides resulted a dose-dependent reduction in raised serum creatinine, β2 -microglobulin, and blood urea nitrogen and a subsequent increase in serum total protein and albumin in nephrotoxic rats (p<.05). An attenuation of the doxorubicin-induced features of renal parenchymal injury was observed on H- and E-stained sections of the kidney tissues. Nootkatone, dehydroaromadendrene, isokhusenic acid, α-vetivone, and isolongifolene were identified in the methanol extract of V. zizanioides based on the GC-MS chromatogram analysis. The findings revealed that the supplement of standardized aqueous root extract of V. zizanioides had a significant dose-dependent nephroprotective activity against doxorubicin-induced experimental nephrotoxicity. PRACTICAL APPLICATIONS: Vetiveria zizanioides is a medicinal plant with a variety of therapeutic applications in kidney-related diseases. Apparently, it is used as a food ingredient due to its fresh and elegant scent and potential bioactivities. The aqueous root extract of V. zizanioides exerted relatively high antioxidant potential in vitro, substantiating the health effects of the plant pertaining to kidney diseases as a potential source of dietary antioxidant. The administration of the plant extract resulted in significant nephroprotection against doxorubicin-induced experimental nephrotoxicity revealing the significance of V. zizanioides as a promising dietary supplement in the management of kidney disease.

Advanced oxidation protein products trigger apoptosis and block epithelial-to-mesenchymal transition in crypt epithelial cells.

Advanced oxidation protein products (AOPPs) are uremic toxins. The present study aimed to investigate the effects of AOPPs on the epithelial mesenchymal transition (EMT) and apoptosis of rat crypt epithelial cells, and to assess the signaling pathways involved. The oxidized rat serum albumin was obtained by sodium hypochlorite modification as AOPPs, and the rat serum albumin (RSA) without sodium hypochlorite modification was set as the control. Different concentrations of AOPPs or RSA were incubated with rat crypt epithelial cells (IEC-6 cells). After culturing for 48 and 72 h, apoptosis was detected by flow cytometry. IEC-6 cells were divided into three groups: A normal group, an AOPPs group and an RSA group. Three groups of cells were collected following treatment for 2 h, and the phosphorylation levels of Akt and p65 NF-κB were detected by western blotting. After 72 h of treatment, the cells were collected and the apoptotic rate was detected by flow cytometry. The expression of EMT-related proteins was detected by reverse transcription-quantitative polymerase chain reaction and western blotting. The apoptotic rate of IEC-6 cells increased with the concentration of AOPPs, and the apoptotic rate of the AOPPs group was higher than that of the RSA group. The expression of fibronectin, snail, slug and collagen I in the AOPPs group was lower than that in the RSA group, while the expression of E-cadherin was not significantly different between the two groups. In addition, the expression of fibronectin, snail, slug and collagen I genes in the AOPPs-treated group was equal to or lower than that in the normal group. Compared with the normal group, the Akt phosphorylation level was decreased and the p65 phosphorylation level was increased in the AOPPs- or RSA-treated groups. Compared with the AOPPs-treated group, Akt and p65 phosphorylation levels in RSA-treated group were slightly higher. In conclusion, AOPPs trigger apoptosis and inhibit the EMT of rat crypt epithelial cells, which may be associated with the inhibition of Akt phosphorylation and the promotion of p65 phosphorylation.

Nutritional, Biochemical and Haematological Indices of White Albino Rats Fed Complementary Diets Developed from Selected Cereals and Legumes

Background: Childhood malnutrition is a common public health problem in Ethiopia. This animal study aims to evaluate the haematological, biochemical, and nutritional indices of weanling albino rats fed on complementary diets developed from selected cereals and legumes. Methods: The nutritional qualities of the formulated blends were assessed biologically by feeding white albino rats in order to determine feed intake, the growth rate, protein quality parameters, biochemical and haematological properties. The commercial weaning diet (cerifam) and casein were used as control diets. On the 28th day, blood samples were collected from three randomly selected rats by decapitation. Full blood count analysis was carried out using the Automated Haematologic Analyzer to determine haematological parameters. All the biochemical parameters were determined using Mindray diagnostic kits. Results and Discussion: The food intake pattern showed that animals placed on diet 3, diet 4, casein, and cerifam consumed more food than those fed on diet 1, diet 2, and diet 7. The mean growth rate of animals fed on diet 3 and diet 4 was significantly (p&lt;0.05) higher than those placed on diet 1, diet 2, and diet 7 but similar to those fed with commercial diet and casein. The protein quality evaluation of the diets showed that the protein efficiency ratio ranged from 1.20 to 2.43 while the biological value was in the range of 54.53 to 69.48%. The net protein utilization and true digestibility were 65.62 to 70.21% and 59.01 to 64.01%, respectively. Serum total protein, albumin, and globulin levels in rats given diet 3 and diet 4 were comparable to the controls and within the normal range. Comparatively, the creatinine and urea levels of rats fed on the control and formulated diets were within the normal range. The serum alanine aminotransferase values of rats fed on the control and formulated diets ranged from 22.03 IU/L in diet 2 to 37.76 IU/L in diet 4 and were not significantly different from each other. In this study, the packed cell volume and haemoglobin values of the control and formulated diets were within the recommended range. Evidently, diet 3 and diet 4 gave the best growth performance after the feeding trials. Conclusion: The haematological and biochemical indices indicate that these diets can support effective growth and development in rats, and consumption of these diets has no detrimental effects on the liver and renal function.

Interaction of SZV 1287, a novel oxime analgesic drug candidate, and its metabolites with serum albumin

SZV 1287 is our novel multi-target analgesic, which seems to be a promising drug candidate for the treatment of neuropathic pain. Therefore, the drug development process has been started in 2016. Since the pharmacokinetic characterization of a drug candidate is essential and albumin binding of drugs can strongly affect their pharmacokinetic properties, herein we provided the detailed investigation and characterization of the interaction of SZV 1287 and its known metabolites with serum albumin. In a preliminary animal study, we demonstrated the appearance of SZV 1287, oxaprozin, L 2799, L 2805, and L 2811 in the circulation after the per os administration of the parent compound to rats. Then albumin-ligand interactions were examined employing fluorescence spectroscopy, affinity chromatography, ultrafiltration, ultracentrifugation, and molecular modeling. Finally, we tested the potential species dependent differences in the albumin binding of SZV 1287, employing human, bovine, porcine, and rat serum albumins. Our results demonstrated that SZV 1287 and its metabolites form highly stable complexes with albumin (Ka = 105 to 106 L/mol). Furthermore, SZV 1287 occupies Sudlow’s Site II on human serum albumin. Therefore, it is reasonable to hypothesize that SZV 1287-albumin interaction is an important issue regarding the pharmacokinetics of this drug candidate.

Characteristics of bilirubin photochemical changes under green light-emitting diodes in humans compared with animal species

Phototherapy using light-emitting diodes (LEDs) centered on the green spectrum, which has a high cyclobilirubin production rate, was as effective as that centered on the blue spectrum for neonatal hyperbilirubinemia. There are no reports of species differences in bilirubin photochemical changes in this spectrum, and the characteristics of bilirubin photochemical changes in humans must be elucidated to proceed with the development of new light sources that include these spectra. This report describes the characteristic photochemical kinetics of bilirubin under green-spectrum LEDs in human, rat, rabbit, dog, pig, sheep, bovine and chicken serum albumin and rhesus monkey serum. These albumin-bilirubin complex solutions were irradiated by green LEDs, and the time-course changes in bilirubin photoisomers were measured by high-performance liquid chromatography. The cyclobilirubin production rates in humans, pigs, and monkeys were significantly higher than those in other species. The rate constant of (EZ)-cyclobilirubin production from (EZ)-bilirubin ‘k’ was significantly higher in humans and monkeys than in other species. In conclusion, bilirubin photochemical kinetics under green spectrum LEDs in humans were characterized by a high cyclobilirubin production rate at a low substrate concentration. The bilirubin photochemical kinetics in monkeys were similar to those in humans.

High level stable expression of recombinant HIV gp120 in glutamine synthetase gene deficient HEK293T cells

Due to the important pathological roles of the HIV-1 gp120, the protein has been intensively used in the research of HIV. However, recombinant gp120 preparation has proven to be difficult because of extremely low expression levels. In order to facilitate gp120 expression, previous methods predominantly involved the replacement of native signal peptide with a heterologous one, resulting in very limited improvement. Currently, preparation of recombinant gp120 with native glycans relies solely on transient expression systems, which are not amendable for large scale production. In this work, we employed a different approach for gp120 expression. Besides replacing the native gp120 signal peptide with that of rat serum albumin and optimizing its codon usage, we generated a stable gp120-expressing cell line in a glutamine synthetase knockout HEK293T cell line that we established for the purpose of amplification of recombinant gene expressions. The combined usage of these techniques dramatically increased gp120 expression levels and yielded a functional product with human cell derived glycan. This method may be applicable to large scale preparation of other viral envelope proteins, such as that of the emerging SARS-CoV-2, or other glycoproteins which require the presence of authentic human glycans.

Advanced Oxidation Protein Products Induce G1/G0-Phase Arrest in Ovarian Granulosa Cells via the ROS-JNK/p38 MAPK-p21 Pathway in Premature Ovarian Insufficiency.

The mechanism underlying the role of oxidative stress and advanced oxidation protein products (AOPPs) in the aetiology of premature ovarian insufficiency (POI) is poorly understood. Here, we investigated the plasma AOPP level in POI patients and the effects of AOPPs on granulosa cells both in vitro and in vivo. KGN cells were treated with different AOPP doses, and cell cycle distribution, intracellular reactive oxygen species (ROS), and protein expression levels were measured. Sprague–Dawley (SD) rats were treated daily with PBS, rat serum albumin, AOPP, or AOPP+ N-acetylcysteine (NAC) for 12 weeks to explore the effect of AOPPs on ovarian function. Plasma AOPP concentrations were significantly higher in both POI and biochemical POI patients than in controls and negatively correlated with anti-Müllerian hormone and the antral follicle count. KGN cells treated with AOPP exhibited G1/G0-phase arrest. AOPP induced G1/G0-phase arrest in KGN cells by activating the ROS-c-Jun N-terminal kinase (JNK)/p38 mitogen-activated protein kinase (MAPK)-p21 pathway. Pretreatment with NAC, SP600125, SB203580, and si-p21 blocked AOPP-induced G1/G0-phase arrest. In SD rats, AOPP treatment increased the proportion of atretic follicles, and NAC attenuated the adverse effects of AOPPs in the ovary. In conclusion, we provide mechanistic evidence that AOPPs may induce cell cycle arrest in granulosa cells via the ROS-JNK/p38 MAPK-p21 pathway and thus may be a novel biomarker of POI.